This study aimed to construct full-length cDNA clones of the Japanese encephalitis virus (JEV). SA14-14-2 strain and discuss the feasibility of constructing chimeric viruses for exogenous gene expression based on the JEV genetic skeleton. Long-fragment RT-PCR techniques were applied to amplify JEV cD-NAs, and two amplified fragments with corresponding restriction endonuclease sites at both ends were cloned into the pACYC184 vector sequentially. Using standard molecular techniques, the enhanced green fluorescent protein (EGFP) gene was inserted into the 3' non-coding region of JEV as a reporter gene. After in vitro transcription and transfection procedures, wild-type JEV and chimeric JEV that expressed the EGFP as the reporter gene were successfully rescued. The recovered viruses were characterized by RT-PCR, plaque assays, and direct fluorescence microscopy. After six serial passage generations, the stability of the recovered viruses were studied in terms of virus growth characteristics and structural gene expression. The results showed that cDNA clones of rJEV and rJEV-EGFP were successfully constructed and rescued in BHK-21 cells after in vitro transcription and transfection. Each generation of the recovered viruses was stable and the chimeric virus rJEV-EGFP could stably express EGFP. The findings of this study indicate that both rJEV and rJEV-EGFP could be constructed and rescued in BHK-21 cells, and the JEV SA14-14-2 strain could be obtained as a viral vector to express foreign genes.
Cloning, Molecular ; DNA, Complementary ; genetics ; metabolism ; Encephalitis Virus, Japanese ; genetics ; metabolism ; Encephalitis, Japanese ; virology ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Vaccines, Attenuated ; genetics ; metabolism ; Viral Vaccines ; genetics ; metabolism

Adult ; Humans ; Imaging, Three-Dimensional ; Jaw Fixation Techniques ; Joint Dislocations ; diagnostic imaging ; etiology ; surgery ; Male ; Mandible ; surgery ; Mandibular Condyle ; diagnostic imaging ; injuries ; surgery ; Mandibular Fractures ; complications ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed

OBJECTIVETo explore the clinical efficacy of the arthroscopic Mender II stapler for the treatment of patients with meniscus anterior horn injury needing meniscal suture repair.

METHODSAmong 47 patients with meniscus anterior horn injury, 29 patients were male and 18 patients were female, ranging in age from 12 to 31 years old, with a mean age of (20.53± 4.12) years old. The duration of disease ranged from 3 to 35 days, and the average duration was (12.43±5.74) days. The Mender II stapler was used to carry out arthroscopic suture from outside to inside. The Lysholm knee scoring system was used to evaluate and analyze preoperative and postoperative symptoms, such as pain, limping embolism and so on.

RESULTSForty-six patients were followed up, and the duration ranged from 12 to 48 months, with a mean of (20.53±4.12) years. The incision healed at the first stage without important vessels and nerves injuries. The symptoms of the locked knee joint disappeared, and symptoms such as pain, limp, swelling and limitation of activity improved. The Lysholm score increased from preoperative 52.33±7.31 to postoperative 86.74±6.92.

CONCLUSIONUsing Mender II stapler to treat patients with meniscus anterior horn injury who were treated with arthroscopic suture from outside to inside is effective to improve symptoms, and to obtain good short-term results.

Adolescent ; Adult ; Arthroscopy ; methods ; Child ; Female ; Humans ; Male ; Menisci, Tibial ; surgery ; Suture Techniques ; Tibial Meniscus Injuries

With patients' general situation, medication use, occurrence time of adverse drug reaction/event (ADR/ADE), clinical manifestations and prognosis as reference items, a retrospective study was made for 315 cases with ADR/ADE induced by Gastrodin in Chongqing from January 2008 to June 2014, in order to analyze the characteristics of ADR/ADE and provide reference for rational clinical medication. The results showed that among the 315 cases with ADR/ADE, 143 cases (45.4%) were males and 172 cases (54.6%) were females, most of them (74.9%) were aged above 45; 60 cases (19.0%) with ADE were caused by off-label indications and 66 cases (21.0%) with ADE were caused by over dosage; ADR/ADE cases induced by intravenous drip mainly happened within 30 min (85.5%), ADR/ADE cases induced by oral administration mainly happened within 2 h (74.4%), and all of ADR/ ADE cases induced by intramuscular injection happened within 10 min. Totally 593 ADR/ADE cases were reported, which were mainly damages in gastrointestinal system, skin and its adnexa; And 61.9% of ADR/ADE cases were newly reported. It is suggested that medical workers shall learn about the regularity and characteristics of ADR/ADE induced by gastrodin, apply it in clinic with standards, pay close attention to changes of patients' situations and attach importance to the monitoring of ADR/ADE, so as to enhance the safety of medication.
Adolescent ; Adult ; Aged ; Benzyl Alcohols ; administration & dosage ; adverse effects ; Child ; Child, Preschool ; China ; epidemiology ; Drug-Related Side Effects and Adverse Reactions ; epidemiology ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Female ; Gastrointestinal Tract ; drug effects ; Glucosides ; administration & dosage ; adverse effects ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Skin ; drug effects ; Young Adult

To investigate the epidemic and evolutionary trends of enterovirus (EV) in the external environment of Yunnan Province, China, molecular typing was performed on 4 EV strains that were isolated from environmental sewage in Yunnan. The VP1 region of isolates was amplified by RT-PCR using universal enterovirus primers, and the amplified VP1 region was sequenced for GenBank BLAST search and genotype analysis. The 4 EV strains were identified as ECHO7. Their nucleotide and amino acid homologies with the VP1 sequences of 68 ECHO7 strains retrieved from GenBank were measured by Mega software analysis. Our findings showed that ECHO7 strains from environmental sewage and population samples were in different evolutionary branches. These strains showed typical geographical and temporal differences; In addition, there were different transmission chains at the same time and in the same area. ECHO7 strains isolated from sewage water and patients with acute flaccid paralysis during the same period in Yunnan belonged to different clusters and evolved at different speeds. Special concerns are needed for this problem. Continuous molecular biological surveillance of human EV in the external environment of Yunnan will provide strong support for early warning of EV diseases.
China ; Databases, Genetic ; Enterovirus ; genetics ; isolation & purification ; Evolution, Molecular ; Humans ; Molecular Epidemiology ; Sequence Analysis ; Sewage ; virology

Eleven type 1 diabetic patients who received fixed regime of insulin Glargine were included in the study.The levels of fasting serum insulin were measured for each subject at 6:00 in three consecutive mornings.The variability of mean fasting serum insulin in each subject was 3.3%-41.5% (mean 15.4%).The variability did not correlate with the dose of Glargine statistically.

Objective To introduce the experience and analyze the clinical outcome of applica- tion of reinforced bar-ring hybrid external fixator combined with limited internal fixation for complex frac- turcs of tibia plateau in patients.Methods From Aug.2002 to Feb.2005,21 patients with complex fractures of tibial plateau were treated with reinforced bar-ring hybrid external fixatar combined with lim- ited internal fixation.According to Schatzker classification,there were 13 patients with typeⅤfracture and 8 with typeⅥfracture.Voluntary and non-weight beating knee joint motion to partially weight bear- ing was taken one week after operation.Progressive loading was taken continuously until there was clinical evidence of fracture stability and then full weight bearing was done.Results The mean follow-up peri- od was 15.9 months (10-24 months).All the fractures got united and the mean union time was 4.2 months(3-8 months).All the injured knees could extend to less than 5 degrees and flex to more than 90 degrees after rehabilitation.Evaluated by Merchant score system for knee joint,81% of patients achieved excellent healing (7 cases) and good healing (10 cases).Conclusion Operation by using reinforced bar-ring hybrid external fixator combined with limited internal fixation can reduce complications and avoid stiffness of the knee joint.It is an effective method for complex fractures of tibial plateau.

This study is aimed at investigating the potentials of ex vivo expansion and pluri-differentiation of cryopreservation of adult human bone marrow mesenchymal stem cells (hMSCs) into chondrocytes, adipocytes and neurocytes. Cryopreserved hMSCs were resuscitated and cultured for 15 passages, and then induced into chondrocytes, adipocytes and neurocytes with corresponding induction medium. The induced cells were observed for morphological properties and detected for expressions of type II collagen, triglyceride or neuron-specific enolase and nestin. The result showed that the resuscitated cells could differentiate into chondrocytes after exposure to transforming growth factor beta(1) (TGF-beta(1)), insulin-like growth factor I (IGF-I) and vitamin C (V(C)), and uniformly changed morphologically from a spindle-like fibroblastic appearance to a polygonal shape in three weeks. The induced cells were heterochromatic to safranin O and expressed cartilage matrix-procollagenal (II) mRNA. The resuscitated cells cultured in induction medium consisting of dexamethasone, 3-isobutyl-1-methylxanthine, indomethacin and IGF-I showed adipogenesis, and lipid vacuoles accumulation was detectable after 21 d. The resuscitated hMSCs were also induced into neurocytes and expressed nestin and neuron specific endolase (NSE) that were special surface markers associated with neural cells at different stage. This study suggested that the resuscitated hMSCs should be still a population of pluripotential cells and that it could be used for establishing an abundant hMSC reservoir for further experiment and treatment of various clinical diseases.
Bone Marrow Cells ; cytology ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cells, Cultured ; Cryopreservation ; methods ; Humans ; Mesenchymal Stromal Cells ; cytology ; Pluripotent Stem Cells ; cytology ; Tissue Engineering ; methods

OBJECTIVETo analyze the prognostic factors in elderly patients with acute myeloid leukemia (AML).

METHODSThe clinical data of 211 AML patients with age 55 years or over and treated in Shanghai Jiaotong University Medical School affiliated Ruijin Hospital from 2007 to 2011 were collected and analyzed. Multivariate and univariate analysis of clinical data were performed using a Cox regression model and log-rank test, including age, subtype, performance status, white blood cell count, serum LDH and albumin level, and treatment strategy.

RESULTSAcute promyelocytic leukemia (APL) patients had longer survival than other subtypes. To rule out the impact of APL on the prognostic analysis, we conducted multivariate and univariate analysis excluding APL patients. The significant parameters of the univariate analysis were age (P = 0.003), achieving remission (P < 0.01), performance status (P < 0.01), organ dysfunction (P < 0.01), increased WBC counts (P = 0.022), increased LDH level (P = 0.006) and low albumin level (P < 0.01). Multivariate analysis showed that only failure of achieving remission (P < 0.01), poor performance status (ECOG 3-4) (P < 0.01) and increased WBC counts (P < 0.01) were independent prognostic factors. The patients aged 70 years or over had poor overall survival, and no significant difference of OS was observed among patients with age between 55 and 69 years. For patients aged 55 - 69 years received either DA/IA or CAG treatments had longer survival than those with palliative treatments. For those aged 70 years or over, only patients with CAG treatment had significantly longer survival than palliative treatment. For the patients with age less than 70 years and achieving complete remission after induction, intermediate-dose cytarabine consolidation might not improve survival.

CONCLUSIONElderly AML patients should be treated individually. The intermediate-dose cytarabine consolidation might not improve survival of elderly AML patients.

Aged ; Aged, 80 and over ; Female ; Humans ; Leukemia, Myeloid, Acute ; diagnosis ; drug therapy ; Male ; Middle Aged ; Survival Rate

OBJECTIVETo study the mechanism of integrin in hypertrophic scar.

METHODSFibroblasts from 10 samples of human hypertrophic scars was cultured, FQ-PCR assay was applied to detect mRNA expression of alpha-SMA in hypertrophic scar fibroblasts after integrin and FAK antibody blocking.

RESULTSmRNA of alpha-SMA in fibroblasts expressed obviously lower after integrin and FAK antibody blocking than that of control groups ( P < 0.05).

CONCLUSIONThrough accelerating the synthesis of alpha-SMA, integrin and FAK play an important role in contracture of hypertrophic scar.

Actins ; biosynthesis ; Adult ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Contracture ; metabolism ; pathology ; Fibroblasts ; metabolism ; Focal Adhesion Kinase 1 ; biosynthesis ; Humans ; Integrins ; biosynthesis ; RNA, Messenger ; metabolism ; Young Adult